Objective To explore the relationship between the inhibitory effect of quercetin on AC16 cardiomyocyte proliferation and the Wnt/β-catenin signaling pathway,the relationship between quercetin(quercetin,QCT)and the proliferation of AC16 cardiomyocytes through in vitro was investigated.Methods AC16 cells were stimulated with different concentrations of QCT.The effects of QCT on AC16 cell proliferation were detected by inverted microscope photography,trypan blue counting,and CCK-8 assay.Western blot was used to detect the effects of QCT on the expression of phosphorylated β-catenin(p-β-catenin),c-Myc,β-catenin,low density lipoprotein receptor-related protein 5(LRP5),and LRP6.The effect of quercetin on cell proliferation was detected after overexpressing β-catenin ΔN,LRP5,and LRP6 genes,and after silencing LRP5 and LRP6 genes by trypan blue counting.Results Compared with the control group,QCT could decrease the number of AC16 cells and inhibit the proliferation rate,which was concentration-dependent.At the protein expression level,10 and 20 μmol/L QCT led to an significant upregulated modification of p-β-catenin protein(P<0.05)and significant downregulation of c-Myc,β-catenin,LRP5,and LRP6 protein expression(P<0.05)in AC16 cells.Therefore,10 μmol/L QCT was chosen as the intervention concentration.Overexpression of β-catenin ΔN,LRP5,and LRP6 genes in AC16 cells significantly rescued the cell proliferation inhibition caused by 10 μmol/L QCT compared to the drug-only group(P<0.05).Conversely,silencing LRP5 and LRP6 genes led to inhibition of AC16 cell proliferation,and the combination with 10 μmol/L QCT did not exacerbate the inhibition(P>0.05).Conclusion The quercetin could inhibit the Wnt/β-catenin signaling pathway via significant downregulation of LRP5/6,thereby attenuate cell proliferation of AC16 cardiomyocytes.

Background and Aims:N6-methyladenosine(m6A)epigenetic modification plays a crucial role in post-transcriptional gene expression regulation and various physiological and pathological processes,including tumorigenesis.The m6A reader IGF2BP2 significantly enhances mRNA stability and translation efficiency and is abnormally expressed in multiple cancers.However,the specific biological function of IGF2BP2 in pancreatic cancer remains unclear.Therefore,this study investigated the expression of the m6A reader IGF2BP2 in pancreatic cancer and its effects on pancreatic cancer cell functions.Methods:The expression levels of m6A-related writers,erasers,and readers were analyzed using The Cancer Genome Atlas(TCGA),the Genotype-Tissue Expression(GTEX)database,and the Gene Expression Omnibus(GEO).Kaplan-Meier survival analysis was conducted to assess the relationship between IGF2BP2 expression and the prognosis of pancreatic cancer patients.Immunohistochemistry was used to validate IGF2BP2 expression in clinical specimens of pancreatic cancer tissues and adjacent normal tissues.Functional experiments,including CCK-8 assay,flow cytometry for cell cycle analysis,colony formation assay,and Transwell migration assay,were performed to evaluate changes in cell proliferation,cell cycle distribution,colony formation ability,and migration capacity after IGF2BP2 knockdown in pancreatic cancer cells.Results:TCGA-GTEX and GEO database analyses showed that IGF2BP2 was highly expressed in pancreatic cancer tissues(both P<0.05)and that its high expression was associated with poor overall survival(both P<0.05).Immunohistochemical staining of clinical specimens confirmed that IGF2BP2 protein expression was higher in pancreatic cancer than in adjacent normal tissue.Functional experiments demonstrated that IGF2BP2 knockdown significantly reduced the proliferation ability of pancreatic cancer cells,arrested more cells in the G0-G1 phase,decreased colony formation,and impaired cell migration(all P<0.05).Conclusion:The m6A reader IGF2BP2 is highly expressed in pancreatic cancer tissues and is closely associated with poor prognosis in patients with this disease.Its mechanism of action may be related to the promotion of cancer cell growth and migration.

Objective To explore the relationship between the inhibitory effect of quercetin on AC16 cardiomyocyte proliferation and the Wnt/β-catenin signaling pathway,the relationship between quercetin(quercetin,QCT)and the proliferation of AC16 cardiomyocytes through in vitro was investigated.Methods AC16 cells were stimulated with different concentrations of QCT.The effects of QCT on AC16 cell proliferation were detected by inverted microscope photography,trypan blue counting,and CCK-8 assay.Western blot was used to detect the effects of QCT on the expression of phosphorylated β-catenin(p-β-catenin),c-Myc,β-catenin,low density lipoprotein receptor-related protein 5(LRP5),and LRP6.The effect of quercetin on cell proliferation was detected after overexpressing β-catenin ΔN,LRP5,and LRP6 genes,and after silencing LRP5 and LRP6 genes by trypan blue counting.Results Compared with the control group,QCT could decrease the number of AC16 cells and inhibit the proliferation rate,which was concentration-dependent.At the protein expression level,10 and 20 μmol/L QCT led to an significant upregulated modification of p-β-catenin protein(P<0.05)and significant downregulation of c-Myc,β-catenin,LRP5,and LRP6 protein expression(P<0.05)in AC16 cells.Therefore,10 μmol/L QCT was chosen as the intervention concentration.Overexpression of β-catenin ΔN,LRP5,and LRP6 genes in AC16 cells significantly rescued the cell proliferation inhibition caused by 10 μmol/L QCT compared to the drug-only group(P<0.05).Conversely,silencing LRP5 and LRP6 genes led to inhibition of AC16 cell proliferation,and the combination with 10 μmol/L QCT did not exacerbate the inhibition(P>0.05).Conclusion The quercetin could inhibit the Wnt/β-catenin signaling pathway via significant downregulation of LRP5/6,thereby attenuate cell proliferation of AC16 cardiomyocytes.

In recent years,organs-on-chips(OOCs)technology has gradually matured and been recognized by regulatory agencies,playing a key role in the construction of pathophysiological models,pharmacodynamic screening,toxicological evaluation and other fields,providing guidance for the research and development of innovative drugs of Chinese materia medica and the clinical application of classic prescriptions.This article reviewed the application progress of OOCs technology in the research of Chinese materia medica from four dimensions:pharmacodynamic and toxicological evaluation,quality control,active component analysis and innovative drug research and development,in order to provide a reference for the precise application of this technology in the field of Chinese materia medica.

Liu Baoyi was a famous febrile disease expert in the late Qing Dynasty,who innovated the theory of latent febrile disease.Liu proposed that the deficiency of kidney essence and the deficiency of healthy qi were the prerequisites for the latent cold e-vil,and the exogenous cold evil was the inducement of the disease.He advocated the treatment of latent febrile disease based on the six meridians,with the two ideas of eliminating evil qi and strengthening healthy qi as the outline,and the general principle of treatment was to relieve heat,nourish yin and expel evil qi.He attached importance to the coordination of medicine and food,made good use of fresh medicine,attached importance to drug processing,emphasized the use of herb processed by herb,and adapted the medicine ac-cording to the syndromes.He preferred to use co-processed products for clinical use.The study of Liu's academic thoughts and medi-cation characteristics in treating latent febrile diseases is of great value in guiding the treatment and medication of various infectious and epidemic diseases with fever characteristics nowadays.

Objective:To evaluate the feasibility and preventive efficacy of a fascia- locking circular continuous suture ostomy technique in reducing parastomal hernia incidence.Methods:This technique was applied to patients undergoing permanent colostomy following radical rectal cancer resection. Surgical steps included: (1) A circular incision was made 1-2 cm medial to the intersection of the lateral margin of the rectus abdominis muscle and the line connecting the umbilicus to the left anterior superior iliac spine. Subcutaneous tissues were dissected vertically to expose the anterior rectus sheath, followed by blunt separation of the rectus abdominis after longitudinal incision of the sheath. The posterior rectus sheath and peritoneum were similarly incised. (2) Eight equidistant interrupted sutures (anchoring knots) were placed through the anterior rectus sheath, partial rectus abdominis, posterior rectus sheath, and peritoneum. (3) The terminal colon was exteriorized, and continuous sutures were applied to secure the anchoring knots and seromuscular layers of the bowel between knots, forming a circular locking mechanism by tying the terminal suture to the initial knot's tail. (3) The skin and seromuscular layers of the bowel margin were intermittently sutured (8-12 stitches) to achieve mucosal eversion.Results:From February to October 2023, 13 patients (11 males, 2 females; age: 67 ± 10 years; BMI: 23.8 ± 4.0 kg/m2) underwent this technique at the Second Affiliated Hospital of Army Medical University. Mean stoma creation time was 15.7 ± 3.0 minutes. During a follow-up of 14.6 ± 3.1 months, physical examinations and abdominal CT scans identified parastomal hernias in 2 male patients at 10 and 7 months postoperatively. Only one patient experienced a Clavien-Dindo grade ≥Ⅲ complication, which resolved with treatment. No stoma-related complications (e.g., infection, stenosis, or prolapse) occurred in any patient.Conclusion:The fascia-locking circular continuous suture ostomy technique is safe and feasible, demonstrating potential efficacy in preventing parastomal hernia following colostomy.

BACKGROUND:The technique of ultrasound processing is widely used for molecular biological analysis.It is of great significance to study the DNA quality of tissue with different storage years under new ultrasonic treatment for further specimen quality control of molecular detection.OBJECTIVE:To explore the effects of different storage durations on DNA quality in specimens with ultrasound processing to investigate the optimal storage time for molecular tests.METHODS:Forty specimens of breast biopsy were collected and paraffin specimens were prepared by ultrasonography.These specimens were divided into four groups based on their storage periods:<1 year,1-3 years,>3-5 years,and>5 years,which contained 10 cases in each group.Paraffin specimens were sliced;each slice was 3 μm thick;10-15 slices were taken,and DNA was extracted.The mass concentration of DNA was examined by Nanophotometer N60 ultra-micro spectrophotometer and Qubit 4.0 fluorometer.The purity of the DNA was analyzed by the ratio of A260/A280.DNA fragment integrity was measured by capillary electrophoresis (Qsep 100) to evaluate the quality of the DNA fragments.RESULTS AND CONCLUSION:The mean values of A260/A280 in the four groups were between 1.8 and 2.0,meeting the requirements of tests,without significant differences.The mean values of DNA mass concentration (Qubit concentration) were 30.39,14.33,2.52,and 1.95 ng/μL,respectively.The mean values of the N/Q were 6.48,14.18,24.56,and 29.86.The mean values of DNA were:5.64,1.76,1.24,and 0.80.The percentage of large DNA fragments averaged 56.08％,17.72％,12.68％,and 7.90％.Moreover,the Ct values of the internal control detected by PCR were 15.32,17.09,18.39,and 21.24.The three other groups exhibited significantly lower DNA concentration,higher N/Q ratios,decreased DNA quality and percentage of large fragments,and increased values of Ct,compared with the group of within 1 year of storage (P<0.05).The experimental results suggested that for novel ultrasound processed biopsy specimens,we should prioritize samples stored within 1 year for molecular testing.Samples stored within 3 years can also meet the requirements of second-generation sequencing and other tests.Samples stored within 5 years can only be attempted to carry out PCR.Samples stored for more than 5 years were not recommended to carry out molecular tests.

Objective:To investigate the treatment strategies for vesicovaginal fistula（VVF）in women following cervical cancer radiotherapy.Methods:A retrospective analysis was performed on the clinical data of 33 female patients with post-radiotherapy VVF after cervical cancer treatment at Shanghai Sixth People’s Hospital between January 2020 and June 2024. The patients were categorized into three groups based on surgical approaches：Group A（11 patients）：Underwent prone-position VVF repair. Mean age：（50.0±9.6）years；mean radiotherapy sessions：（22.6±2.2）. All had simple VVF without concurrent intestinal or surrounding soft-tissue fistulas. Among them，1 patient previously received laparoscopic VVF repair，1 transvaginal VVF repair，and 2 gracilis muscle flap packing for VVF repair. One month prior to surgery，the average daily usage of urine pads was 16.7（12.8，25.7）pieces，and the quality of life（QOL）score stood at 4.0（4.0，5.0）points. Preoperative cystoscopy revealed that 8 cases had fistulas located in the trigonal region of the bladder，while 3 cases had fistulas on the posterior bladder wall. The diameter of the fistula openings，measured under a microscope，was 1.2（0.8，1.6）cm. Based on the Goh classification criteria for vesicovaginal fistulas，this group included 3 cases of type 1，4 cases of type 2，2 cases of type 3，and 2 cases of type 4；7 cases of type A and 4 cases of type B；as well as 3 cases of typeⅠ，7 cases of type Ⅱ，and 1 case of type Ⅲ.Group B（20 patients）：Underwent gracilis muscle flap packing for VVF repair. Mean age：（58.6±8.8）years；mean radiotherapy sessions：（29.8±3.9）. Three patients had concurrent rectovaginal fistulas and received colostomy for fecal diversion. History of previous interventions：3 had laparoscopic VVF repair，4 transvaginal VVF repair，and 1 both transvaginal and laparoscopic VVF repair. One month prior to surgery，the average daily usage of urine pads was 19.7（15.8，27.7）pieces，and the QOL score stood at 5.0（5.0，6.0）points. Preoperative cystoscopy revealed that 13 cases had fistulas located in the trigonal region of the bladder，while 7 cases had fistulas on the posterior bladder wall. The diameter of the fistula openings，measured under a microscope，was 1.8（1.0，3.2）cm. Based on the Goh classification criteria for vesicovaginal fistulas，this group included 4 cases of type 1，9 cases of type 2，3 cases of type 3，and 4 cases of type 4；6 cases of type A，11 cases of type B and 3 cases of type C；as well as 1 cases of type Ⅱ，and 19 case of type Ⅲ. Group C（2 patients）：Underwent ureterocutaneous diversion. Ages：67 and 73 years；radiotherapy sessions：51 and 60，respectively. Both had concurrent rectovaginal fistulas and bladder soft-tissue fistulas. The patient presented with recurrent thigh abscesses accompanied by fever. One month prior to surgery，the daily usage of urine pads was 29 and 23 pieces，respectively，and the QOL score was 6 points. Cystoscopic examination revealed that the vesicovaginal fistulas were located in the trigone of the bladder，with diameters of 3 cm and 4 cm，respectively. Additionally，partial defects were noted in the ventral wall of the urethra，while no bladder soft tissue fistulas were detected. According to the Goh classification for vesicovaginal fistulas，both cases were categorized as type 4，type C，and type Ⅲ. For Groups A and B，urinary catheters were indwelled for 3 weeks postoperatively，then removed to assess spontaneous urination and incontinence. QOL was evaluated，with a minimum 6-month follow-up. For confirmed postoperative VVF recurrence，re-repair was performed 3?6 months later based on patient preference. For Group C，double-J stents were placed in the ureters，and stoma bags were applied 3 days postoperatively. Stents were replaced every 1?2 months，with QOL assessment. Successful fistula repair in Groups A and B was defined as the absence of vaginal leakage confirmed by cystoscopy after six months of the procedure with no vaginal leakage. For Group C，surgical success was determined by the resolution of perineal urinary leakage and improvement in QOL.Results:All 33 patients completed surgery successfully. Group A：Follow-up duration：16.3（9.6，24.6）months. Surgical repair succeeded in 7 patients，with unobstructed spontaneous urination and no vaginal incontinence. Four patients had VVF recurrence：2 refused further treatment，and 2 underwent repeat gracilis muscle flap packing. One was successfully repaired，while one recurrence case refused further treatment. Group B：Follow-up duration：17.0（9.5，24.8）months（8?32 months）. Thirteen patients restored spontaneous urination without recurrence. Seven had recurrence：5 refused further surgery，and 2 underwent re-repair. One repair succeeded without incontinence，while one recurrence case refused treatment. Group C：Follow-up durations were 6 and 22 months. Perineum remained dry without incontinence（no urine pads needed），and no recurrence of thigh soft-tissue redness/infection occurred. QOL scores were 2 and 3，respectively.Conclusions:Post-radiotherapy VVF in women after cervical cancer presents complex and variable conditions. The primary goal of treatment should be to improve patients’ quality of life. Treatment approaches should be selected based on the complexity of urinary fistulas and local tissue conditions. In general，patients who are younger，have received lower doses of radiation therapy，present with smaller fistula diameters，have well-vascularized and elastic perifistular tissues，and have no concurrent tissue fistulas are candidates for prone-position VVF repair. Patients who do not meet the criteria for transvaginal repair，have a history of at least two previous repair attempts，or have concurrent vaginorectal fistulas require gracilis muscle flap packing for VVF repair. Patients with three or more types of concurrent tissue fistulas，extensive pale and inelastic perifistular tissues，and who are not amenable to repair surgery undergo ureterocutaneous diversion.

Turning to critical illness is a common stage of various diseases and injuries before death. Patients usually have complex health conditions, while the treatment process involves a wide range of content, along with high requirements for doctor′s professionalism and multi-specialty teamwork, as well as a great demand for time-sensitive treatments. However, this is not matched with critical care professionals and the current state of medical care in China. Telemedicine, which shortens the distance of medical professionals and the gap of disease diagnosis and treatments in various regions through electronic information, can effectively solve the current problem. Therefore, there is an urgent need to develop a standardized, high-quality visualization telemedicine round system .Therefore, experts have been organized to search domestic and foreign literature on telemedicine round for critically ill patients and to form this consensus based on clinical experiences so as to further improve the level of critical care treatments in regions.

In recent years,organs-on-chips(OOCs)technology has gradually matured and been recognized by regulatory agencies,playing a key role in the construction of pathophysiological models,pharmacodynamic screening,toxicological evaluation and other fields,providing guidance for the research and development of innovative drugs of Chinese materia medica and the clinical application of classic prescriptions.This article reviewed the application progress of OOCs technology in the research of Chinese materia medica from four dimensions:pharmacodynamic and toxicological evaluation,quality control,active component analysis and innovative drug research and development,in order to provide a reference for the precise application of this technology in the field of Chinese materia medica.